Histochemical tracing of lysosomal enzymes. Improved preparation technique for acid phosphatase, beta-glucuronidase, acid-beta-galactosidase, and arylesterase in rat liver.
The efficiency of the membrane methods of Meijer (1972) and Lojda (1973) in histochemical lysosome tracing was studied and compared with the results of a preparation technique developed by us, employing fresh-frozen celloidin-coated sections; in addition, these methods were compared with results obtained with conventional formalin-sucrose-fixed livers. The lysosomes were traced by reactions for ACPase, beta-glucuronidase, arylesterase and acid-beta-galactosidase activity in rat livers systematically harvested at different time points of occurence of their maximal and minimal activities during the 24-h period, thus indicating a heterogeneity of lysosomes. 3. Optimal results with respect to the morphological appearance of lysosomes were obtained in fresh-frozen celloidin coated sections. This preparation method also caused no enzyme inhibition or loss, thus delivering comparatively the strongest reactions in livers and in enzyme models. 4. Day-time-dependent extralysosomal enzyme activities regularly occur in hepatocytes. Extralysosomal localizations however, are not a consequence of technically induced enzyme diffusion; they are best visualized in celloidin-coated sections; the membrane method produces less satisfactory results, and formalin-sucrose-fixed livers were least satisfactory.
D000135:Acid Phosphatase / Q000378:metabolism; D000818:Animals; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism; D002940:Circadian Rhythm; D005966:Glucuronidase / Q000378:metabolism; D006651:Histocytochemistry / Q000379:methods*; D006867:Hydrolases / Q000378:metabolism*; D008099:Liver / Q000201:enzymology*; D008247:Lysosomes / Q000201:enzymology*; D008297:Male; D051381:Rats; D011917:Rats, Inbred Lew; D013048:Specimen Handling / Q000379:methods; D001616:beta-Galactosidase / Q000378:metabolism
D006867:Hydrolases; D002265:Carboxylic Ester Hydrolases; C019734:arylesterase; D000135:Acid Phosphatase; D001616:beta-Galactosidase; D005966:Glucuronidase